IL-4R Recombinant Rabbit Monoclonal Antibody [PSH10-86] - BSA and Azide free

Western blot analysis of IL-4R on different lysates with Rabbit anti-IL-4R antibody (HA751378) at 1/2,000 dilution.

Lane 1: THP-1 cell lysate (20 µg/Lane)
Lane 2: Daudi cell lysate (20 µg/Lane)
Lane 3: 293T cell lysate (20 µg/Lane)
Lane 4: K-562 cell lysate (20 µg/Lane)
Lane 5: Jurkat cell lysate (20 µg/Lane)
Lane 6: MCF7 cell lysate (20 µg/Lane)
Lane 7: Ramos cell lysate (20 µg/Lane)
Lane 8: RAW264.7 cell lysate (20 µg/Lane)
Lane 9: PC-12 cell lysate (20 µg/Lane)
Lane 10: Mouse spleen tissue lysate (40 µg/Lane)
Lane 11: Mouse pancreas tissue lysate (40 µg/Lane)

Predicted band size: 90 kDa
Observed band size: 110 kDa

Exposure time: 1 minute; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751378) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

IL-4R was immunoprecipitated from 0.2 mg Jurkat cell lysate with HA751378 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751378 at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: Jurkat cell lysate (input)
Lane 2: HA751378 IP in Jurkat cell lysate
Lane 3: Rabbit IgG instead of HA751378 in Jurkat cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 51 seconds; ECL: K1801

Immunocytochemistry analysis of PC-12 cells labeling IL-4R with Rabbit anti-IL-4R antibody (HA751378) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-IL-4R antibody (HA751378) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.